Immunomodulatory drug lenalidomide is a synthetic compound derived by modifying the chemical structure of thalidomide to improve its potency and reduce its side effects. Lenalidomide is a 4-amino-glutamyl analogue of thalidomide that has emerged as a drug with activity against various hematological and solid malignancies. It is approved by FDA in USA for clinical use in myelodysplastic syndromes with deletion of chromosome 5q and multiple myeloma. Studies have shown that lenalidomide exert anti-tumor activity probably by various mechanisms in hematologic malignancies, such as immunomodulation, anti-angiogenesis and effects on signal transduction. In this article, the progresses of study on these problems are reviewed.
Hematologic Neoplasms ; immunology ; Humans ; Immunologic Factors ; Thalidomide ; analogs & derivatives ; pharmacology

To investigate the influence of As2O3, dexamethasone (Dex) and thalidomide (Thal) on apoptosis-induced myeloma cell line U266 cytoplasmic calcium concentrations ([Ca2+]i), U266 cells were incubated in the culture of RPMI 1640 with 15% FBS in 24-well plate and exposed to different concentrations of As2O3, Dex and Thal for 8 hours, respectively, then cell apoptosis was analyzed by fluorescence microscopy and flow cytometry (FCM) with Annexin V-FITC/PI double staining, and cytoplasmic free calcium were detected on FCM through Fluo-3/AM loading. The results indicated that (1) apoptotic cells were gradually increased with enhancement of As2O3, Dex and Thal concentrations; (2) apoptotic cell rates increased from 0.56% in control to 31.54%, 28.35% and 21.97% respectively after treatment with As2O3, Dex and Thal; (3) As2O3, Dex induced U266 cell apoptosis accompanied with raise of [Ca2+]i; (4) [Ca2+]i had no statistically significant changes in Thal-induced apoptotic U266 cells. It is concluded that the raise of [Ca2+]i is one of the mechanisms for As2O3 and Dex-induced U266 cells apoptosis, whereas Thal-induced U266 apoptosis has no significant relation to [Ca2+]i changes.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Calcium ; metabolism ; Cell Line, Tumor ; Cytoplasm ; metabolism ; Dexamethasone ; pharmacology ; Humans ; Multiple Myeloma ; pathology ; Oxides ; pharmacology ; Thalidomide ; pharmacology

Animals ; Cell Adhesion Molecules ; metabolism ; Liver Cirrhosis, Experimental ; drug therapy ; metabolism ; Male ; Rats ; Rats, Wistar ; Thalidomide ; pharmacology ; therapeutic use

Lenalidomide, a novel immunomodulatory agent, is a kind of thalidomide derivatives, which shows a good efficacy and safety for hematological system diseases. This review is aimed to evaluate the efficacy and safety of lenalidomide in treatment of patients with multiple myeloma, chronic lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, classical Hodgkin's lymphoma and POEMS syndrome at their replased or refractory state. At the same time, this review focuses on the newest clinical research and the latest application progress of lenalidomide for relapsed or refractory hematological system diseases.
Antineoplastic Combined Chemotherapy Protocols ; Humans ; Lenalidomide ; Leukemia, Lymphocytic, Chronic, B-Cell ; Lymphoma, Non-Hodgkin ; Multiple Myeloma ; Thalidomide ; analogs & derivatives ; pharmacology

BACKGROUNDGlioma is the most common primary brain tumor with poor prognosis. Temozolomide has been used with thalidomide to treat gliomas. We investigated the synergistic mechanism of these two drugs in vitro.

METHODSHuman malignant glioma cells U251-MG were cultured and assigned to four groups with different treatments for 3 days: temozolomide group (100 micromol/L), thalidomide group (100 microg/L), temozolomide (100 micromol/L) plus thalidomide group (100 microg/L) and control group. MTT assay was applied to evaluate the cell viability. Cell cycle was analyzed by flow cytometry. The ultra-structural features of autophagosomes were observed with electron microscope. Acridine orange and monodansylcadaverine were adopted to label autophagosomes and flow cytometry was applied for quantification of autophagosomes. The expression of autophagy-associated protein was detected by Western blotting.

RESULTSProliferation of tumor cell was obviously suppressed by temozolomide with thalidomide treatment than by either drug used alone (P = 0.000 for each day). The combination treatment induced cell cycle arrest at G0/G1 phase. Typical autophagic ultra-structural character was found after the combined treatment. Thalidomide promoted the autophagy induced by temozolomide. The autophagy-associated proteins-microtubule associated protein 1 light chain 3 (MAP1LC3) and Beclin1 were more significantly up-regulated by the combined treatment than temozolomide used alone (MAP1LC3, P = 0.000; Beclin1, P = 0.004). The expression level of phosphatase and tensin homolog deleted on chromosome ten (PTEN), which promoted autophagy by suppressing PI3K/Akt/mTOR signaling pathway, was elevated by thalidomide (thalidomide group: P = 0.000; combined group: P = 0.002).

CONCLUSIONSThalidomide enhances the cytotoxicity of temozolomide by promoting the autophagy induced by temozolomide. Contributing to the up-regulation of PTEN by thalidomide, the expression of autophagy associated protein-MAP1LC3 and Beclin1 was enhanced, which leads to a reinforced autophagy in the combined treatment of temozolomide and thalidomide in vitro.

Antineoplastic Agents, Alkylating ; pharmacology ; Autophagy ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dacarbazine ; analogs & derivatives ; pharmacology ; Glioma ; pathology ; Humans ; Thalidomide ; pharmacology

OBJECTIVETo explore the inhibitory effect of thalidomide combined with interferon (IFN) on the human acute myeloid leukemia cell line Kasumi- 1 and its mechanism.

METHODSThe inhibitiory effect of Kasumi- 1 cells by thalidomide, interferon or combination was detected by CCK- 8 method, the apoptosis by flow cytometry, the expression of apoptosis related proteins by Western blot, vascular endothelial growth factor (VEGF) concentration in culture supernatant by ELISA.

RESULTSThalidomide inhibited the proliferation of Kasumi- 1 in a dose- dependent manner from 50 μg/ml to 500 μg/ml with an IC₅₀ of (451.13 ± 6.92）μg/ml at 24 h and (362.50 ± 14.52）μg/ml at 48 h. IFN also demonstrated the inhibitory capacity in a dose-dependent manner from 500 U/ml to 5 000 U/ml, with an IC₅₀ of (2 209 ± 127) U/ml at 24 h and (1 393±63) U/ml at 48 h. The apoptosis rates of Kasumi-1 cells treated with thalidomide 350 μg/ml or IFN 1 400 U/ml for 48 h were (14.68 ± 2.61) % and (21.71 ± 0.71)%, respectively, significantly higher than control group (P<0.01). In combination group the inhibition and the apoptosis rate were (88.50 ± 2.40) % and (41.95 ± 3.41)%, significantly higher than control and each single agent group (P<0.01). The VEGF concentrations of combination group [(94.61 ± 5.46) ng/L decreased significantly, as compared to thalidomide group [(141.11 ± 3.70) ng/L and IFN group [(119.90 ± 2.00) ng/L (P < 0.05). Western blot analysis showed Bcl-2 expression of Kasumi-1 cells decreased, while p-P38, Bax, cytochrome C, cleaved-Caspase-3, 8, 9 increased after treated with thalidomide 350 μg/ml or IFN 1 400 U/ml for 48 h. When treated with the combination agents, the expression of Bcl-2 further decreased and p-P38, Bax, cytochrome C, cleaved-Caspase-3, 8, 9 further increased as compared with each single agent (P < 0.05).

CONCLUSIONThalidomide and IFN could synergistically inhibit the proliferation of Kasumi-1 cells probably through inducing apoptosis via the mitochondrial pathway, death receptor pathway and P38 MAPK pathway, as well as inhibiting VEGF secretion.

Apoptosis ; Caspases ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Interferons ; pharmacology ; Leukemia, Myeloid, Acute ; pathology ; MAP Kinase Signaling System ; Thalidomide ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism

To investigate the influence of the thalidomide on the growth of multiple myeloma cells from untreated, relapsed or refractory patients and summarize its mechanisms, thalidomide influence on colony growth of untreated, relapsed or refractory multiple myeloma cells cultured by semisolid methylcellulose was observed. The level of interleukin-6 (IL-6) autosecreted by myeloma cells was tested by IL-6-dependent cell line when myeloma cells were treated with thalidomide at 200 microgram/ml, and in the same concentration of thalidomide the expression of IL-6 receptor were tested by flow cytometry. Results showed that colony growths of myeloma cell from untreated and relapsed or refractory patients were all colonies were inhibited when treated by thalidomide up to 75 microgram/ml or 100 microgram/ml concentration. The inhibition was concentration-dependent, higher concentration cause more inhibition. After treatment with thalidomide at 200 microgram/ml, the concentrations of IL-6 secreted by myeloma cells were (148.5 +/- 96.7) microgram/ml, and the levels of IL-6 receptor expressed on the cell surface were 16.7% and 20.2% in untreated and relapsed or refractory patients, respectively, and those were significantly lower than those levels in the cells before exposure to thalidomide. It was concluded that thalidomide can inhibit growth of both relapsed or refractory cells and untreated myeloma cells in vitro. Therefore, it can be used to treat untreated multiple myeloma patients. Inhibiting tumor cells secreting level of IL-6 and reducing the expression of IL-6 receptor on myeloma cell surface is one of the mechanisms for thalidomide to remedy multiple myeloma patients
Angiogenesis Inhibitors ; pharmacology ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Interleukin-6 ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Receptors, Interleukin-6 ; biosynthesis ; Thalidomide ; pharmacology ; Tumor Cells, Cultured

Apoptosis ; drug effects ; Cell Line, Tumor ; Drug Synergism ; Flavanones ; administration & dosage ; pharmacology ; Humans ; Multiple Myeloma ; pathology ; Thalidomide ; administration & dosage ; analogs & derivatives ; pharmacology

The study was aimed to investigate the anti-myeloma molecular mechanism of thalidomide (TLD) by detecting gene expression profiles of human myeloma cell line RPMI8226 treated with thalidomide. cDNA microarray were used to detect thousands of gene expression in gene chip. Two cDNA probes were prepared through reverse transcription from mRNA of RPMI8226 cells untreated and treated with TLD. These two probes were labeled with Cy3 and Cy5 fluorescence dyes respectively, then hybridized with cDNA microarray containing 1152 different human genes. The genes with differential expression in RPMI8226 cells treated with TLD for 72 hours were screened by scanning and analysis of computer software, and their functions were explored. The results showed that after co-culture of RPMI 8226 cells with TLD in 100 micromol/L concentration for 72 hours, 22 genes with differential expression were screened. Among these genes, the expressions of 4 genes were down-regulated including rpl32 gene, scya3 gene, mmp1 gene and igbp1 gene. Eighteen genes were up-regulated including wars gene, tubb4q gene, ube1l gene, txnrd1 gene and fyb gene. The study indicated that (1) wars gene encoding tryptophanyl-tRNA synthetase was up-regulated by TLD, while mmp1 gene encoding matrix metalloprotein 1 was down-regulated, they may be related to the inhibition of angiogenesis caused by TLD. (2) scya3 gene encoding macrophage inflammatory protein-1alpha and igbp1 gene encoding immunoglobulin binding protein 1 were down-regulated by TLD, they may play a role in the inhibition of cell proliferation caused by TLD. (3) tubb4q gene encoding tubulin beta4, ube1l gene encoding ubiquitin-activating enzyme E1-like protein and txnrd1 gene encoding thioredoxin reductase 1 were up-regulated by TLD, they may involve in apoptosis of RPMI8226 cells induced by TLD. (4) fyb gene encoding Fyn-binding protein was up regulated by TLD which associated with killing MM cells. It is concluded that 22 differentially expressed genes are involved in protein synthesis and degradation, cell signal transduction, cytoskeletal movement, immune modulation, cell metabolism, regulation of anti-oncogene and cell apoptosis, which relate directly or indirectly to molecular mechanisms of anti-myeloma effects induced by TLD.
Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Multiple Myeloma ; genetics ; Oligonucleotide Array Sequence Analysis ; Thalidomide ; pharmacology

OBJECTIVETo investigate the effect of thalidomide on Annexin II (AnxA2) gene regulation in multiple myeloma cell line RPMI8226 and human microvascular endothelial cell line HMEC-1 cells in vitro, and explore the potential mechanism of thrombosis induced by thalidomide.

METHODSRPMI8226 and HMEC-1 cells were cultivated in vitro. Real time quantitative PCR (RQ-PCR) was used to detect the influence of thalidomide at different concentration on the expression of AnxA2 mRNA, flow cytometry (FCM) and confocal microscopy were used to detect the cell surface protein level after the samples were stimulated with different concentrations of thalidomide.

RESULTSAnxA2 mRNA level in RPMI8226 cells treated with thalidomide at 12.5 microg/ml, 25.0 microg/ml and 50.0 microg/ml was decreased compared with the control group (0.60+/-0.15, 0.33+/-0.14, 0.42+/-0.16, vs 1.07+/-0.16, respectively, P<0.05) and did so in HMEC-1 cells (0.21+/-0.20, 0.08+/-0.08, 0.17+/-0.16 vs 1.16+/-0.24, respectively, P<0.05). The AnxA2 protein level in RPMI8226 cells treated with above mentioned concentrations of thalidomide was also decreased compared with the control (3.39+/-0.32, 2.82+/-0.28, 3.21+/-0.23 vs 5.53+/-0.32, respectively, P<0.05) and that did so in HMEC-1 cells (0.72+/-0.11, 0.64+/-0.08, 0.67+/-0.08 vs 1.40+/-0.15, respectively, P<0.05).

CONCLUSIONSThalidomide can inhibit the expression of AnxA2 mRNA and protein in RPMI8226 and HMEC-1 cells, which may be one of the mechanisms for the development of thrombosis induced by thalidomide in multiple myeloma patients.

Annexin A2 ; genetics ; metabolism ; Cell Line ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; Humans ; Multiple Myeloma ; metabolism ; pathology ; RNA, Messenger ; genetics ; Thalidomide ; pharmacology